WO2019088034A1 - Dispositif de réglage de zone d'observation, dispositif de commande d'imagerie, procédé de fonctionnement de dispositif de réglage de zone d'observation et programme de réglage de zone d'observation - Google Patents

Dispositif de réglage de zone d'observation, dispositif de commande d'imagerie, procédé de fonctionnement de dispositif de réglage de zone d'observation et programme de réglage de zone d'observation Download PDF

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Publication number
WO2019088034A1
WO2019088034A1 PCT/JP2018/040156 JP2018040156W WO2019088034A1 WO 2019088034 A1 WO2019088034 A1 WO 2019088034A1 JP 2018040156 W JP2018040156 W JP 2018040156W WO 2019088034 A1 WO2019088034 A1 WO 2019088034A1
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WIPO (PCT)
Prior art keywords
observation area
observation
area
overlapping
container
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PCT/JP2018/040156
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English (en)
Japanese (ja)
Inventor
佑介 和多田
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富士フイルム株式会社
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Priority to JP2019550377A priority Critical patent/JPWO2019088034A1/ja
Publication of WO2019088034A1 publication Critical patent/WO2019088034A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/26—Stages; Adjusting means therefor

Definitions

  • the present invention relates to a technique for setting each observation area in tiling imaging.
  • Pluripotent stem cells such as ES (Embryonic Stem) cells and iPS (Induced Pluripotent Stem) cells have the ability to differentiate into cells of various tissues, and they can be used in regenerative medicine, drug development, disease elucidation, etc. It is noted that it can be applied in
  • pluripotent stem cells such as ES cells and iPS cells or cells induced to differentiate are imaged with a microscope or the like and the characteristics of the image are captured to evaluate the differentiation state of the cells etc. .
  • each observation area in the well is scanned by moving the stage on which the well plate or the like is installed with respect to the imaging optical system to photograph each observation area, and then imaging for each observation area
  • a method has been proposed for joining images to generate a composite image.
  • Patent Document 1 discloses a method in which the position of the bottom surface of the culture vessel is measured in advance, and the bottom surface of the culture vessel is automatically focused and photographed.
  • the overlapping area that is, the overlapping area
  • the number of captured images may increase when all the observation objects are captured, which may result in an increase in the amount of data and a longer imaging time.
  • the present invention has been made in view of the above-mentioned circumstances, and by appropriately setting the overlapping degree of overlapping areas in adjacent observation areas, it is possible to prevent an increase in the amount of data of photographed images and an increase in photographing time. To aim.
  • the observation area setting device overlaps a part of a new observation area adjacent to the observation area set in advance with respect to the observation area set in advance with respect to the observation object stored in the container.
  • An observation area setting unit that sequentially sets new observation areas; An overlapping degree in which the overlapping degree of the overlapping overlapping area is set larger as the inclination degree of the bottom surface of the container positioned in the previously set observation area in the alignment direction of the previously set observation area and the new observation area is larger.
  • a setting unit that sequentially sets new observation areas; An overlapping degree in which the overlapping degree of the overlapping overlapping area is set larger as the inclination degree of the bottom surface of the container positioned in the previously set observation area in the alignment direction of the previously set observation area and the new observation area is larger.
  • the “previously set observation area” refers to an area set before “new observation area” is set, and “first” is a time-series relationship. means.
  • the “arrangement direction” refers to a direction along a line connecting the centers of the previously set observation area and the new observation area.
  • the observation area setting device of the present invention includes a shape information receiving unit that receives shape information of the bottom surface of the container,
  • the overlap degree setting unit may acquire the inclination degree based on the shape information input from the shape information reception unit.
  • the inclination degree may be a value based on a difference between the heights of both ends in the arranging direction of the bottom surface of the container located in the observation area set in advance.
  • the inclination degree is a size including the one side on which the new observation area is set in the observation area set in advance and the size in agreement with the observation area set in advance. It may be a value based on the difference in height between both ends in the alignment direction of the bottom surface of the containers located in the area of height.
  • the observation area may be smaller than the container.
  • the container may be a dish, a well plate or a flask.
  • An imaging control apparatus is the observation area setting apparatus described above; And a control unit configured to cause the imaging unit to image the observation target stored in the container for each observation region set by the observation region setting device.
  • the imaging control apparatus of this invention may be provided with the shape information storage part which memorize
  • the shape information storage unit may store a table in which identification information of the container is associated with shape information of the bottom surface of the container.
  • the operation method of the observation area setting device of the present invention is An operation method of an observation area setting device comprising an observation area setting unit and an overlapping degree setting unit,
  • the observation area setting unit newly overlaps a part of a new observation area adjacent to the observation area set in advance with respect to the observation area set in advance with respect to the observation object stored in the container.
  • Set various observation areas sequentially
  • the degree of overlapping of the overlapping area is determined by the overlapping degree setting unit.
  • the degree of inclination of the bottom surface of the container located in the previously set observation area in the alignment direction of the previously set observation area and the new observation area is large. Set as large as possible.
  • the method of operating the observation region setting device according to the present invention may be provided as a program that causes a computer to execute the method.
  • Another observation area setting apparatus is a memory for storing an instruction to be executed by a computer.
  • a processor configured to execute the stored instructions, the processor
  • a new observation area is sequentially set by overlapping a part of a new observation area adjacent to the previously set observation area with respect to the previously set observation area with respect to the observation target accommodated in the container
  • the process of setting the overlapping degree of overlapping overlapping areas to be larger as the inclination degree of the bottom surface of the container located in the previously set observation area in the alignment direction of the previously set observation area and the new observation area is larger Run.
  • the overlapping degree of the overlapping area is determined by the inclination of the bottom surface of the container located in the observation area set in the direction in which the observation area and the observation area set in advance are aligned.
  • FIG. 7 shows schematic structure of one Embodiment of the microscope observation system to which the image processing apparatus of this invention is applied.
  • Diagram showing the scanning locus of each observation area in the well plate Figure for explaining the shape information of the bottom of the culture vessel
  • Schematic block diagram showing the configuration of the observation area setting unit
  • a diagram for explaining the overlapping area for each observation area in the culture vessel A diagram showing an example of an acquired captured image
  • Diagram showing multiple overlapping areas set for the culture vessel
  • a partial enlarged view of the observation image at the upper left corner in FIG. 7 A partial enlarged view of the observation image in the upper right corner in FIG. 7
  • Diagram explaining another method of calculating the degree of inclination Flow chart showing processing performed in the present embodiment
  • FIG. 1 is a view showing a schematic configuration of a microscope observation system to which an observation area setting device according to an embodiment of the present invention is applied.
  • the microscope observation system of the present embodiment includes a microscope apparatus 1, a microscope control apparatus 2, an input apparatus 3, and a display apparatus 4.
  • the microscope control device 2 corresponds to the imaging control device of the present invention.
  • the microscope apparatus 1 is a phase-contrast microscope, and for example, images a phase-contrast image of cultured cells as a photographed image as an observation target.
  • the microscope apparatus 1 includes an illumination light irradiation unit 10, an imaging optical system 30, a stage 61, and an imaging unit 40.
  • a culture vessel 60 in which an observation target S such as a cell and a culture solution C are accommodated is installed. At the center of the stage 61, a rectangular opening is formed.
  • the culture vessel 60 is installed on a member forming the opening, and a photographed image of the observation target S in the culture vessel 60 is configured to pass through the opening.
  • a cultured cell group (cell colony) is disposed as the observation target S.
  • the cultured cells include pluripotent stem cells such as iPS cells and ES cells, nerves derived from stem cells, skin, cells of myocardium and liver, skins removed from human body, retina, myocardium, blood cells, nerves and There are cells of organs etc.
  • a well plate having a plurality of wells (corresponding to the container of the present invention) is used, but not limited to this, a petri dish, a flask, a dish or the like may be used.
  • a well plate in which a plurality of wells are arranged is used as the culture vessel 60.
  • the bottom surface in the culture vessel 60 is the installation surface P1 of the observation target S, and the observation target S is disposed on the installation surface P1.
  • the culture solution C is filled in the culture vessel 60.
  • the cells cultured in the culture solution are the observation target S.
  • the observation target S is not limited to those in the culture solution, but water, formalin, ethanol, methanol, etc.
  • the cells fixed in the liquid may be the observation target S.
  • the illumination light irradiation unit 10 irradiates illumination light for so-called phase difference measurement to the observation target S accommodated in the culture vessel 60 on the stage 61, and in the present embodiment, the phase difference A ring-shaped illumination light is emitted as illumination light for measurement.
  • the illumination light irradiator 10 of the present embodiment has a white light source 11 for emitting white light for phase difference measurement, a ring-shaped slit, and the white light emitted from the white light source 11 is incident thereon.
  • the slit plate 12 emits ring-shaped illumination light
  • the condenser lens 13 irradiates the ring-shaped illumination light emitted from the slit plate 12 with respect to the observation target S.
  • the slit plate 12 is provided with a ring-shaped slit for transmitting white light to a light shielding plate for shielding white light emitted from the white light source 11, and the white light passes through the slit to form a ring shape. Illumination light is formed.
  • the condenser lens 13 converges the ring-shaped illumination light emitted from the slit plate 12 toward the observation target S.
  • the imaging optical system 30 forms an image of the observation target S in the culture vessel 60 on the imaging unit 40, and includes an objective lens 31, a phase plate 32, and an imaging lens 33.
  • the phase plate 32 is formed by forming a phase ring on a transparent plate transparent to the wavelength of the ring-shaped illumination light.
  • the size of the slit of the slit plate 12 described above is in a conjugate relationship with this phase ring.
  • phase film for shifting the phase of the incident light by 1 ⁇ 4 wavelength and a light reducing filter for reducing the incident light are formed in a ring shape.
  • the direct light incident on the phase plate 32 passes through the phase ring, so that the phase is shifted by 1 ⁇ 4 wavelength and its brightness is weakened.
  • most of the diffracted light diffracted by the observation target S passes through the portion of the transparent plate of the phase plate 32, and its phase and brightness do not change.
  • the imaging lens 33 receives the direct light and the diffracted light that have passed through the phase plate 32, and images these lights on the imaging unit 40.
  • the imaging unit 40 includes an imaging element that receives an image of the observation target S formed by the imaging lens 33, captures an image of the observation target S, and outputs a phase difference image as an observation image.
  • an imaging element a CCD (charge-coupled device) image sensor, a CMOS (Complementary Metal-Oxide Semiconductor) image sensor, or the like can be used.
  • the stage 61 is driven by the drive unit 62 and moves in the horizontal direction in the X direction and the Y direction orthogonal to each other.
  • each observation area smaller than the wells in each well of the well plate is scanned, and a photographed image for each observation area is acquired by the imaging unit 40.
  • each observation area is scanned by overlapping a part of adjacent observation areas. The overlapping area of adjacent observation areas will be described in detail later.
  • the photographed image for each observation area is output to the microscope control device 2.
  • FIG. 2 is a diagram showing a scanning locus of each observation area by a solid line 77 in the case of using a well plate 70 having six wells 71. As shown in FIG. As shown in FIG. 2, each observation area in the well plate 70 is scanned along the solid line 77 from the scanning start point 75 to the scanning end point 76 by the movement of the stage 61 in the X and Y directions.
  • the photographed image of each observation area in the well is acquired by moving the stage 61
  • the invention is not limited thereto, and the imaging optical system 30 is moved with respect to the stage 61.
  • the photographed image of each observation area may be acquired.
  • both the stage 61 and the imaging optical system 30 may be moved.
  • this invention may scan not only with this but another scanning locus
  • the microscope control device 2 is configured of a computer provided with a CPU (Central Processing Unit) 20, a primary storage unit 24, a secondary storage unit 25, an external I / F (Interface) 28, and the like.
  • the CPU 20 includes a control unit 21, an observation area setting device 22, and an image processing unit 23, and controls the entire microscope observation system.
  • the primary storage unit 24 is a volatile memory used as a work area or the like when executing various programs.
  • An example of the primary storage unit 24 is a RAM (Random Access Memory).
  • the secondary storage unit 25 is a non-volatile memory in which various programs, various parameters and the like are stored in advance, and the shape information 26 which is an example of the shape information storage unit of the present invention is stored.
  • the observation area setting program 27 of the present invention is installed.
  • the observation area setting program 27 When the observation area setting program 27 is executed by the CPU 20, the observation area setting device 22 functions.
  • Examples of the secondary storage unit 25 include an EEPROM (Electrically Erasable Programmable Read-Only Memory), a flash memory, and the like.
  • the external I / F 28 controls transmission and reception of various information between the microscope apparatus 1 and the microscope control apparatus 2.
  • the CPU 20, the primary storage unit 24, and the secondary storage unit 25 are connected to the bus line 29.
  • the external I / F 28 is also connected to the bus line 29.
  • the observation area setting program 27 is distributed by being recorded on a recording medium such as a digital versatile disc (DVD) and a compact disc read only memory (CD-ROM), and is installed in the computer from the recording medium.
  • a recording medium such as a digital versatile disc (DVD) and a compact disc read only memory (CD-ROM)
  • the observation area setting program 27 is stored in a state accessible from the outside with respect to the storage device or network storage of the server computer connected to the network, and after being downloaded to the computer in response to the request from the outside. It may be installed.
  • the shape information 26 is shape information of the bottom of the culture vessel 60, ie, the installation surface P1.
  • the shape information of the bottom surface of the culture vessel 60 is measured in advance using, for example, a laser displacement meter.
  • FIG. 3 is a view for explaining the shape information of the bottom of the culture vessel 60.
  • the shape information of the present embodiment is information obtained by measuring the bottom surface 60a of the culture container 60 with a spatial resolution of 10 ⁇ m ⁇ 10 ⁇ m in the XY direction, as shown in FIG.
  • the spatial resolution of the shape information is not limited to this.
  • the method of measuring the shape information of the bottom surface 60a of the culture vessel 60 is not limited to measurement by a laser displacement meter, and measurement may be performed using other methods such as a confocal method and a spectral interference method.
  • the shape information of the bottom surface 60s of the culture vessel 60 may be measured by a laser displacement meter provided in the microscope device 1 when the culture vessel 60 is installed on the stage 61, or the culture vessel 60.
  • a table may be stored in the secondary storage unit 25 in which the identification information of the object and the shape information measured in advance are associated with each other. Then, the user sets and inputs identification information of the culture vessel 60 installed on the stage 61 using the input device 3, and reads out the shape information of the culture vessel 60 having the identification information from the secondary storage unit 25. It is also good.
  • a barcode or the like may be given to the culture container 60 and the identification information may be acquired by reading the barcode, instead of the user performing setting input.
  • the identification information of the culture vessel 60 may be, for example, a model number of a manufacturer or a serial number.
  • the dedicated computer may be firmware that executes a program stored in a non-volatile memory, such as a built-in ROM (Read Only Memory) or a flash memory.
  • a dedicated circuit such as an application specific integrated circuit (ASIC) or field programmable gate arrays (FPGA) that permanently stores a program for executing at least a part of functions of the microscope control device 2.
  • ASIC application specific integrated circuit
  • FPGA field programmable gate arrays
  • the program instruction stored in the dedicated circuit may be combined with the program instruction executed by the general-purpose CPU programmed to use the program of the dedicated circuit.
  • the computer hardware configuration may be combined to execute program instructions.
  • the control unit 21 controls the drive of the illumination light irradiation unit 10, the drive unit 62 that drives the stage 61, the imaging optical system 30, and the imaging unit 40, and acquires a captured image of the observation target S.
  • the control unit 21 also functions as a display control unit that causes the display device 4 to display one composite phase difference image generated by combining the phase difference images of the respective observation positions captured by the microscope device 1. .
  • the control unit 21 also causes the imaging unit 40 to image the observation target S.
  • the culture vessel 60 is a well plate in which a plurality of wells are arranged, the control unit 21 causes the imaging unit 40 to image each observation region in each well.
  • the observation area setting device 22 sets an observation area for the culture vessel 60.
  • FIG. 4 is a schematic block diagram showing the configuration of the observation area setting device 22.
  • the observation area setting device 22 includes an observation area setting unit 50, an overlap degree setting unit 51, and a shape information reception unit 52.
  • the shape information receiving unit 52 reads and acquires the shape information of the bottom surface of the culture container 60 from the shape information 26 stored in the secondary storage unit 25.
  • the observation area setting device 22 according to the present embodiment includes the shape information receiving unit 52, the present invention is not limited to this, and the shape information receiving unit 52 may not be provided. In this case, the shape information 26 is read from the secondary storage unit 25 as required by an instruction from the CPU 20.
  • the observation area setting unit 50 newly overlaps a part of a new observation area adjacent to the observation area with respect to the observation area S set in advance with respect to the observation target S accommodated in the culture vessel 60.
  • the observation area is set sequentially.
  • FIGS. 5A and 5B illustrate an overlapping area for each observation area in the culture container 60
  • FIGS. 6A and 6B illustrate an example of a captured image acquired.
  • the culture container 60 is a well plate in which a plurality of wells are arranged, an observation region is set in each well.
  • both ends of the bottom of the well are referred to as end A and end B, respectively, and the center of the bottom is referred to as center O.
  • the photographed image photographed by the photographing unit 40 is as shown in FIG.
  • the peripheral portions on the side a and the side b become blurred images.
  • the photographing unit 40 is made to photograph a plurality of photographed images including the overlapping areas. In the overlapping area, it is possible to select a photographed image with better image quality.
  • FIG. 7 shows a diagram showing a plurality of overlapping regions set for the culture vessel 60.
  • each observation area is scanned while overlapping a part of the adjacent observation areas.
  • each observation area Gi i is The number of observation areas
  • the observation areas G1 to G5 are arranged in order from the left end of the upper line (first line), G6 to G12 in order from the right end of the second line, and G13 in order from the left end of the third line.
  • G21 to G21, G22 to G30 from the right end of the fourth line, G31 to G39 from the left end of the fifth line, G40 to G48 from the right end of the sixth line, G49 to G57, eighth line from the left end of the seventh line G58 to G64 are set in order from the right end of G, and G65 to G69 are set in order from the left end of the ninth line.
  • the widths d of the overlapping regions are all shown to have the same size in the drawing, but in reality, the overlapping degree setting unit 51 sets the widths d to different sizes.
  • FIG. 8 is an enlarged view of the observation image at the upper left corner in FIG. 7, and FIG. 9 is an enlarged view of the observation image at the upper right corner in FIG.
  • the overlapping area between the observation area G1 at the upper left corner in FIG. 7 and the observation area G2 adjacent to the right is an overlap area K1, the observation area G2 and the observation area G3 adjacent to the right
  • the observation region G1 is the previously set observation region of the present invention
  • the observation region G2 is a new observation region with respect to the observation region G1
  • the observation region G2 is the previously set observation region
  • the observation area G3 becomes a new observation area with respect to the observation area G2.
  • the overlap degree setting unit 51 sets an overlap area of the overlap area, that is, the overlap area K1 and the overlap area K2 in FIG.
  • the overlapping area is set at a ratio of, for example, 20 to 30% with respect to one observation area. From the shape information of the bottom of the culture vessel 60 (in the present embodiment, the well 71) located in the observation area G1, the overlapping degree setting unit 51 first arranges the observation area G1 and the observation area G2 in the arrangement direction, that is, in FIG. The height of the bottom surface corresponding to the center position of the sides of the observation area G1 in the direction of the arrow M1 (X direction) is acquired, and the difference in height between the two ends is calculated.
  • the value of the difference in height is taken as the inclination degree of the overlapping area K1.
  • the height of the bottom corresponding to the center position of the sides at both ends of the observation area G1 has been acquired, but not limited to this, the average value of the heights at the positions corresponding to the sides at both ends is determined
  • the difference between the average heights may be taken as the difference between the heights at both ends.
  • the inclination degree is not limited to the use of the value of the height difference itself at both ends of the observation area, and may be newly set based on the value of the height difference in the overlapping area K1, for example.
  • the inclination degree of the overlapping area K2 is also calculated in the same manner as the overlapping area K1. That is, the heights of both ends in the direction of the arrow M1 of the observation area G2 are acquired, and the difference between the heights of the both ends is calculated to be the inclination degree.
  • the bottom surface of the culture container 60 (well 71) of the present embodiment has the shape shown in FIG. 5, the direction of the arrow M1 inclines the bottom surface further from the center of the culture container 60. Is larger, the calculated inclination degree is such that the overlapping area K1 is larger than the overlapping area K2.
  • the overlap degree setting unit 51 sets the width d2 of the overlap area K2 smaller than the width d1 of the overlap area K1 so that the overlap area K1 is larger than the overlap area K2.
  • widths d1 to d4 of overlapping regions K1 to K4 shown in FIG. 7 are set.
  • the initial width, ie, the width d1 of the overlapping area K1 is determined in advance so that the overlapping area K1 is set at a ratio of, for example, 20 to 30% with respect to the observation area G1. It is also possible to read out the initial setting value.
  • a correspondence table in which the relationship between the height difference and the width d is defined may be stored in the secondary storage unit 25 and the width d1 may be set based on the correspondence table.
  • the overlapping degree setting unit 51 sets the overlapping degree of the overlapping area K5 and the overlapping area K6 in FIG. Based on the shape information of the bottom of the culture vessel 60 (in the present embodiment, the well 71) located in the observation area G5, the overlapping degree setting unit 51 first arranges the observation area G5 and the observation area G6 in the arrangement direction, that is, in FIG. The height of the bottom surface corresponding to the center position of the sides of the both ends of the observation area G5 in the direction of the arrow M2 (Y direction) is acquired, and the difference in height between the two ends is calculated. In this embodiment, the value of the height difference is taken as the inclination degree of the overlapping area K5.
  • the inclination degree of the overlapping area K6 is also calculated in the same manner as the overlapping area K5. That is, the height of the bottom surface corresponding to the center position of the sides of the observation area G6 in the direction of the arrow M2 of the observation area G6 is acquired, and the difference in height between the two ends is calculated to be the inclination degree.
  • the direction of the arrow M2 inclines the bottom of the culture vessel 60 further from the center thereof. Is larger, the calculated inclination degree is larger in the overlapping area K5 than in the overlapping area K6.
  • the overlap degree setting unit 51 sets the width d6 of the overlap area K6 smaller than the width d5 of the overlap area K5 so that the overlap area K5 is larger than the overlap area K6.
  • the overlap area from the overlap area K8 between the observation area G7 and the observation area G8 to the overlap area K11 between the observation area K10 and the observation area K11 is The same widths as the widths d1 to d4 of the overlapping regions K1 to K4 shown in FIG. 7 are set.
  • the width d7 of the overlap area K7 between the observation area G6 and the observation area G7 and the width d12 of the overlap area K12 between the observation area G11 and the observation area G12 are set in the same manner as the above method. As described above, the width d of the overlapping area K is set for each column and row in the direction of the arrow M1 (including the direction of the arrow M3) and the direction of the arrow M2 in FIG.
  • the width d of the overlapping area K is set for each column and row for each of the direction of arrow M1 (including the one for scanning the direction of arrow M3) and the direction of arrow M2 in FIG.
  • the invention is not limited to this, and the width d of the overlapping area K may be set for each column or row, or overlapping may be performed in the direction of the arrow M1 and the direction of the arrow M2 for each observation area.
  • the width d of the area K may be set.
  • FIG. 10 shows a diagram for explaining another method of calculating the degree of inclination.
  • the overlap degree setting unit 51 includes the culture vessel 60 located in a region Gh including the side h on the side of setting the observation region G2 of the observation region G1 and having the same size as the observation region G1.
  • the height of the bottom surface corresponding to the center position of the sides of the area Gh in the alignment direction of the observation area G1 and the observation area G2 is acquired from the shape information of the bottom of the well 71).
  • the difference in height of the bottom surface corresponding to the center position may be calculated as the inclination degree.
  • the position of the area Gh can be appropriately changed by the user.
  • the observation area setting device 22 is configured, and the observation area setting unit 50 observes the observation target S housed in the culture vessel 60 based on the overlapping degree set by the overlapping degree setting unit 51. Set the area sequentially.
  • the degree of overlapping of overlapping areas in adjacent observation areas can be suitably set according to the shape of the culture vessel 60, so that increase in the data amount of photographed images and prolongation of photographing time can be prevented. can do.
  • the image processing unit 23 performs various processing such as gamma correction, luminance / color difference conversion, and compression processing on the image signal acquired by the imaging unit 16. Further, the image processing unit 23 outputs an image signal obtained by performing various processes to the control unit 21 for each frame at a specific frame rate. Further, the image processing unit 23 generates a single composite image by combining the phase difference images of the respective observation areas captured by the microscope device 1. As for the overlapping area, the image of the overlapping part is selected to have a better image quality. For example, a photographed image with the highest contrast may be used as an image of the overlapping area, or an overlapping portion of the photographed image closest to the center of the culture container 60 may be used as an image of the overlapping area.
  • the overlapping region may be set large. However, if the overlapping image is set to a large size, the number of captured images increases, and it takes time to capture images. As in the present invention, by setting the overlapping degree to be larger as the inclination of the container is larger, it is possible to maintain the high image quality of the composite image and to prevent an increase in the data amount of the captured image and an increase in the imaging time.
  • the input device 3 includes a mouse, a keyboard, and the like, and receives various setting inputs by the user.
  • the display device 4 displays the composite image generated by the image processing unit 53, and includes, for example, a liquid crystal display. Further, the display device 4 may be configured by a touch panel and used as the input device 3.
  • FIG. 11 is a flowchart showing the process performed in the present embodiment.
  • the shape information receiving unit 52 acquires shape information of the bottom surface of the culture container 60 (step S1).
  • the overlapping degree setting unit 51 calculates the difference in height of the bottom surface of the culture vessel 60 located in the observation region set earlier, ie, the observation region G1 in FIG. 8 from the shape information acquired in step S1 ( Step S2) This difference in height is acquired as the degree of inclination (step S3).
  • the overlap degree setting unit 51 determines the overlap degree for each observation area as described above (step S4), and the observation area setting unit 50 sequentially selects the observation areas based on the overlap degree determined by the overlap degree setting unit 51.
  • the setting is made (step S5), and the process is ended.
  • the observation area setting device 22 sets the observation area as described above.
  • the control unit 21 causes the imaging unit 40 to capture an image for each observation area set by the observation area setting device 22, acquires captured images for each of a plurality of observation areas, and a plurality of image processing units 23 Are combined to generate a composite image Gs. Then, the generated composite image Gs is displayed on the display device 4 and provided for observation.
  • the present invention is not limited to a phase contrast microscope, and can be applied to other microscopes such as a differential interference microscope and a bright field microscope. .
  • the overlapping degree of the overlapping areas in the adjacent observation areas can be suitably set according to the shape of the container, so that the increase in the data amount of the photographed image and the prolongation of the photographing time are prevented. be able to.

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Abstract

La présente invention évite une augmentation du volume de données d'une image photographiée et une prolongation de la durée de photographie dans : un dispositif de réglage de zone d'observation ; un dispositif de commande d'imagerie ; un procédé pour faire fonctionner un dispositif de réglage de zone d'observation ; et un programme de réglage de zone d'observation. Lorsque de nouvelles zones d'observation doivent être définies séquentiellement pour un objet d'observation logé dans un récipient en formant un chevauchement partiel entre une zone d'observation précédemment définie et une nouvelle zone d'observation adjacente à la zone d'observation précédemment définie, le degré de chevauchement dans la région de chevauchement est défini de manière à être plus élevé lorsque le niveau d'inclinaison de la surface inférieure du récipient situé dans la zone d'observation précédemment définie, dans une direction dans laquelle la zone d'observation précédemment définie et la nouvelle zone d'observation sont alignées, est plus élevé.
PCT/JP2018/040156 2017-11-06 2018-10-29 Dispositif de réglage de zone d'observation, dispositif de commande d'imagerie, procédé de fonctionnement de dispositif de réglage de zone d'observation et programme de réglage de zone d'observation WO2019088034A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013201530A (ja) * 2012-03-23 2013-10-03 Canon Inc 撮像装置及びその制御方法
JP2016173511A (ja) * 2015-03-17 2016-09-29 キヤノン株式会社 画像取得装置、及びそれを用いた画像取得方法
JP2017161385A (ja) * 2016-03-10 2017-09-14 株式会社Ｓｃｒｅｅｎホールディングス 撮像装置における撮像配置決定方法および撮像装置

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013201530A (ja) * 2012-03-23 2013-10-03 Canon Inc 撮像装置及びその制御方法
JP2016173511A (ja) * 2015-03-17 2016-09-29 キヤノン株式会社 画像取得装置、及びそれを用いた画像取得方法
JP2017161385A (ja) * 2016-03-10 2017-09-14 株式会社Ｓｃｒｅｅｎホールディングス 撮像装置における撮像配置決定方法および撮像装置

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